16-alkylene pregnene derivatives



United States Patent 3,369,272 16-ALKYLENE PREGNENE DERIVATIVES DavidTaub, Metuchen, Norman L. Wendler, Summit,

and Robert D. Holfsommer, Jr., Metuchen, N.J., as-

signors to Merck & Co., Inc., Railway, N.J., a corporation of New JerseyNo Drawing. Filed Apr. 24, 1961, Ser. No. 104,319

6 Claims. (Cl. 167-65) CHgOR' wherein R is alkylene, R is hydrogen oracyl, R" is hyd-roxy or oxygen, and X is hydrogen or halogen. The brokenline between carbon atoms 1 and 2 indicates that a double bond may bepresent in this position.

The 16-alkylene steroids pnoduced in accordance with the presentinvention possess extremely high anti-inflammatory activity,considerably greater than that of the parent steroids, and areespecially effective for the treatment of arthritis and related diseasessince they can be administere-d for their cortisone-like action in lowdosage thereby minimizing undesired side effects.

In preparing our novel chemical compounds, the starting materialutilized may be a I i-substituted-17uhydroxy-1l-oxygenated-16-alkylene-20-pregnanoneor 3-substituted17a-hydroxy-1l-oxygenated-l6-alkylene-20-allopregnanoe which may beidentified by the following structural formula:

wherein X is hydrogen or halogen, Y is alpha or beta hydrogen, Z isoxygen, dioxolane, alpha or beta acyloxy or hydroxy, R is alkylene, andR" is hydroxy or oxygen as above, with or without a double bond at the 5(6)-position.

In a preferred embodiment of our invention, 16-alkyl- 3,309,272 PatentedMar. 14, 1967 ene-3u,17a-dihydroxy-11,20-pregnanedione which isrepresented by the following formula:

(IJHs O:

is utilized as the starting material. However, it is clear to thoseskilled in the art that other starting materials, such as thosedescribed above, may be similarly converted to the desired end products.

It has been found that l6-alkylene-3x,17a-dihydroxy- 11,20-pregnanedionemay be caused to react with bromine in a chloroform medium to form16-alkylene-21-bromo- 3u,17ot-dihydroxy-11,20-pregnanedione which hasthe following formula:

CHiBr wherein R is as above.

Treatment of the above16-alkylene-2l-bromo-3a,17adihydroxy-l1,20-pregnanedione with potassiumiodide, potassium acetate and acetic acid in acetone yields 16-alkylene304,17on 21 trihydroxy-l1,20-pre'gnane dione-Zlacetate which may berepresented graphically as follows:

(llH OAc 0o --0H Oxidation of 16-alkylene-3a,17u,21-trihydroXy-11,20-pregnanedione 21-acetate with sodium dichromate results in the formationof 16-a1ky1ene-17a,2l-dihydroxy-S,11,20- pregnanetrione 21-acetate whichhas the following structural formula:

CHaOAo wherein R is as above.

Treatment of 16-alkylene-17a,21-dihydroxy-3,11,20 pregnanetrione21-acetate with 1 mole of bromine yields 3 1-6 alkylene4-bromo-l7tx,2l-dihydroxy-3,l1,20-pregnanetrione which has the followingstructural formula:

C I-IzOAc wherein R is as above Reaction of the abovel6-alkylene-4-bromo-l70;,21-bromo-17a,2l-dihydroxy-3,l1,20-pregnanetrione 2l-acetate withsemicanbazide hydrohalide results in the formation of the3-sernica-rbazone of l6-alkylene-17a,2l-dihydroXy-4pregnene-3J1,2O-trione 21-acetate which may be represented by thefollowing formula:

NHzCONHN:

wherein R is as above.

Upon reacting the latter semicarbazone of l6-alkylene-17a,2l-dihydroXy-4-pregnene-3,l1,20-trione with aqueous aceticacid-pyruvic acid there is formed l6-alkylene-17a,2l-dihydroxy-4-pregnene-3,11,2-0-trione ZI-acetate which has thefollowing structure:

CHzOAO wherein R is as above.

The Zl-acetate is hydrolyzed to the alcohol by treatment of1-6-alkylene-17a,21-dihydroxy-4-pregnene-3,11, 20-tr-ione Zl-acetatewith potassium bicarbonate in aqueous methanol to yieldl6-alkylene-l7a,21-dihydroxy-4- pregnene-3,l1,2(]-trione which may berepresented by the following structural formula:

wherein R is as above.

The latter compound is then reacted with semi-cabbazide hydrochloride inmethanol-dimethylformamide to form the 3,20-disemicarbazone of16-alkylene-17a, 21-

dihydroxy-4pregnene-3,11,20-trione which has the following structuralformula:

C=NNHCONH2 as WNQU wherein R is alkylene as above.

Reduction of the 3,20-disemicar-bazone of l6-alkylene-17u,21-dihydroxy-4-pregnene-3,11,20-trione with sodium boronhydrideproduces the 3,20-disemicarbazone of 16- al'kylene 11B,17a,21trihydroxy-4-pregnene-3,20-dione, which has the fol-lowing formula:

wherein R is as above.

The 3,20-disemicarbazone of l6-a1kylene-11B,l7a,21-trihydroxy-4-pregnene-3,ZO-dione is reacted with pyruvic acid in aqueousacetic acid to remove the semicarbazone groups at the 3 and 20-positionsto form 16-alkylene- 11,8,170:,21-trihydroxy-4-pregnene-3,ZO-dione whichmay be chemically represented as follows:

OH OH wherein R is as above.

Other 21-acylates such as the propionate, t-butyl acetate or benzoatemay be prepared by reacting the 21- alcohol with the appropriate acylanhydrides.

The 16 alkylene-l7a,2l-dihydroXy-3,11,20-pregnanetrione 21-acetate maybe reacted with 2 molecules of bromine to form16-alkylene-2,4-dibromo-17a,2l-d.ihy-

droxy-pregnane-3,11,20-trione ZI-acetate which may be represented by thefollowing formula:

wherein R is as above.

Treatment of the above compound with dimethyl aniline indimethylformamide results in the desired 16-alkylene17a,2l-dihydroxy-l,4-pregnadiene-3,11,20-trione 21- acetate representedby the following formula:

wherein R is as above. The 21-acetate can be readily converted to thecorresponding 21-01 by hydrolysis, for example, in aqueous methanolicpotassium bicarbonate to give 16alkylene-17a,21-dihydroxy-1,4-pregnadiene- 3,11,20-trione represented bythe following formula wherein R is as above.

CHZAC C=O oH wherein R is as above and X is hydrogen or fluorine and R"is hydroxy or oxygen.

Treatment of these species with dimethyl aniline in 6 170:,21 dihydroxy11-oxygenated-1,4-pregnadiene-3,20- dione 21-acetate and16-alkylene-9a-fluoro-17a,21-dihydroxy1l-oxygenated-1,4-pregnadiene-3,ZO-dione 21-acetate which may berepresented graphically as follows:

ornoAc wherein R, R" and X are as above. The species represented by theabove formula are highly potent antiinflammatory agents.

The 21-acetate esters of these compounds can be hydrolyzed to thecorresponding alcohols, 16-alkylene-17a, 21 d'ihydroxy 11oxygenated-1,4-pregnadiene-3,20- dione and 16 alkylene9u-fluoro-17a,21-d-ihydroXy-11- oxygenated 1,4 pregnadiene-3,20-dionerepresented by the following structural formula:

wherein R and R" are as above.

The 16-a1kylene compounds described above may also be obtained fromeither the corresponding 16cc or 16;?- alkyl compounds of the followingformula:

in which R", X are as above and R"" is alkyl.

Treatment of this compound with semicarbazide base and se-micarbazidehydrochloride leads to the corresponding 3,20-disemicarbazone of thefollowing formula:

CHzOH C=NNHCONIh wherein R", X and R"" are as above.

The latter 3,20-disemicarbazone on heating in acetic dimethylformamideresults in the desired 16-alkyleneacid and acetic anhydride is convertedinto the corresponding 16-unsaturated 3,20-disemioarbazone of thefollowing formula:

CH2OR I C=NNHCONH2 l li s N HZCONHN wherein R, R", X and R"" are asabove.

The latter 16-unsaturated semicarbazone on treatment with hot aqueousacetic acid is converted to the corresponding 3,20-dione of thefollowing formula:

CHzO R i C O i I!!! fl i wherein R, R", X and R"" are as above.

The latter compound on treatment with alkaline hydrogen peroxide or analkyl hydroperoxide in the presence of a base or an organic peracid isconverted into the corresponding 16a,l7a-oxide of the following formula:

CHzOR CI-IzOR' wherein R, R, R" and X are as above.

The 21-acetates may be converted to the corresponding 21-alcohols of thefollowing formula by acid or basic hydrolysis.

R, R and X are as above.

Other ZI-acylates e.g. the propionate, t-butyl acetate, benzoate may beprepared by reacting the 21-alcohols with the appropriate acylanhydride. Also the 21-esters with inorganic acids e.g. the 21-sulfate,2l-phosph-ate may be prepared in the appropriate manner.

This process is especially useful in preparing 9oc-flll010-11,8,17a,2l-trihydroxy-16-methylene 1,4 pregnadiene 3,20-dione,9oc-fluoro-11B,17a,21-trihydroxy-16-mcthylene- 4-pregnene-3,20-dione andl6-methylene prednisolone.

Alternatively, the 1,4-pregnadiene compounds described above may beprepared from the corresponding 4-pregnene-3-ones by dehydrogenationchemically or microbially by means of Bacillus sphaericus.

These 16-methylene-pregnenes and 16-methylene-pregnadienes mayconveniently be converted to the corresponding lea-methyl and l6B-mcthylcompounds by hydrogenation of the 16-methylene compound in the presenceof a hydrogenation catalyst. When these compounds contain unsaturatedbonds in the A ring, however, these bonds are likewise reduced alongwith the 16-methylene group to produce the corresponding 16-methylpregnane compound, whereupon the desired unsaturation at the C1 and/ orC4 position must be re-introduced either chemically ormicrobiologically. Alternatively, the saturated compound, as for example16-alkylene-l7a,2l-dihydroxy- 3,11,20-pregnane-trione 21-acetate, may behydrogenated directly to form a mixture of the corresponding tmethyl andlop-methyl pregnanes.

The catalytic reduction of the 16-methylene steroid compound ispreferably carried out with palladium-oncalcium carbonate as thecatalyst. The steroid is dissolved in an inert solvent, as for example,ethyl acetate, methanol, ethanol, benzene and reacted with hydrogen at apressure ranging from 1 atmosphere to about 15 atmospheres and at atemperature of from about 10 C. to 50 C. until 1 molar equivalent ofhydrogen is taken up. Preferably the reaction is conducted at 1atmosphere and at a temperature of 25 C., in which case the uptake ofhydrogen is complete in about 1 hour. The reaction mixture is filteredand dried, and the isomers separated each from the other by fractionalcrystallization, chromatography or the like. In carrying out thisreaction, either the ZI-free alcohol or its corresponding 21-ester maybe employed as the starting material.

The resulting lfia-methyl and IGB-mcthyl pregnane compounds may then bebrominated at the 2- and/or 4- carbon atoms followed bydehydrobromination to introduce the desired unsaturation in the A ring.Alternatively, the 16-methyl pregnane compound may be contacted with thedehydrogenating activity of Bacillus sphaericus to produce thecorresponding 1,4-pregnadiene compound.

The starting material for the present process, namely, 30,17os dihydroxy11 oxygenated-16-alkylene-20-pregnanone or the 3-acylate thereof can beprepared from the known 3at-hydroxy-lo-pregnene-l1,20-dione 3-acetate inaccordance with the following procedure:

It has been found that 3a-hydroxy-16-pregne11e-11,20- dione 3-acetatemay be caused to react with diazoalkanes, such diazomethane anddiazoethane to form 1602,1706- alkyleneazo 30c hydroxy-l1,20-pregnanedione B-acetate which has the following structural formula:

wherein R is hydrogen or alkyl.

Upon heating 16a,17u-alkyleneazo-3a-hydroxy-l1,20- pregnanedione3-acetate, there is formed primarily 16- 9alkyl-3u-hydroxy-16-pregnene-11,20-dione 3-acetate which may berepresented by the following formula:

wherein R is alkyl as above Treatment of16-alkyl-3a-hydroxy-l6-pregnene-l1,20- dione S-acetate, with anoxidizing agent such as hydrogen peroxide in the presence of sodiumhydroxide or peracids such as peroxytrifluoracetic acid results in theformation of 165 alkyl-16a,17a-epoxy-3a-hydroxy-11,20-pregnanedionewhich has the structural formula:

OHa

wherein R"" is as above.

Room temperature acetylation gives 16;3-alkyl-l6a,17aepoxy3oc-l1YdIOXY-l 1,20 pregnanedione S-acetate. Upon treatment withhydrogen bromide in acetic acid there is formed 16 alkylene3a,17a,-dihydroxy-11,20-pregnanedione 3-acetate which may be representedas follows:

wherein R is as above.

Hydrolysis of this latter compound in aqueous methanolic potassiumhydroxide leads to 16a1kylene-3a,17adihydroxy-l1,20-pregnanedionerepresented by the following formula:

no j terials. The composition may take the form of tablets, powders,capsules, elixers, syrups or other dosage forms which are particularlyuseful for oral ingestion. Liquid diluents are employed in sterilecondition for par enteral use, namely, by injection. Such a medium maybe a sterile solvent for the active material. For topical administrationany of the known dermatologic vehicles may be employed. Such vehiclesfor external application include water-soluble ointment bases, oils,petrolatum and jellies.

The following experimental part illustrates in detail some of thecompounds which constitute this invention and methods for theirproduction. However, this inven tion is not to be construed as limitedthereby in spirit or in scope since it will be apparent to those skilledin the art that many modifications in materials and methods may be madewithout departing from the invention.

Example 1. Preparation of 21 bromo 304,17adihydroxy-16-methylene-1l,ZO-pregnanedione 3-acetate To a stirredsolution of 209 mg. of 30,l7oL-dlhYdIOXY-16-methylene-l1,20-pregnanedione 3-acetate (prepared as describedhereinbelow) in 4 ml. of chloroform maintained at 4045 C., is slowlyadded mg. of bromine in 5 ml. of chloroform. Uptake of bromine iscomplete within 30 minutes. Ether is added and the mixture washed twicewith cold 5% potassium bicarbonate solution. The organic extract isdried over magnesium sulfate and the colorless residue crystallized fromacetoneether gives colorless prisms of2l-bromo-3u,l7ot-dihydroxy-16-methylene-11,20-pregnanedione MP. 195- 200dec.

Analysis.Calculated for C H O Br: C, 59.88; H, 6.91; Br, 16.60. Found:C, 60.50; H, 6.85; Br, 15.86.

The 30:,17oc hydroxy 16 methylene 11,20- pregnanedione 3-acetate used asa starting material in this example, is prepared starting with the known30chydroxy-16-pregnene-l1,20-dione 3-acetate, in accordance with thefollowing procedure:

In a 500 ml. 3-neck flask equipped with condenser, dropping funnel andnitrogen inlet are placed 20 g. of potassium hydroxide in ml. of water,ml. of methanol and 100 ml. of ether. A solution of 10 g. ofN-methyl-N-nitrosotosylamide in 50 ml. of ether is placed in thedropping funnel.

Diazomethane is generated by warming the generation flask to 404-5 C.and cautiously adding the N- methyl-N-nitrosotosylamide-ether from thedropping funnel. Nitrogen is utilized to sweep the diazomethane into asolution of 20 g. of 3a-hydroxy-16-pregnene-11, ZO-dione 3-acetate in100 ml. of tetrahydrofuran and ml. ether. The process is continued untilthe steroid solution remains yellow for several hours. The product, 30;hydroxy 1601,1704 methylene azo 11,20 pregnanedione 3-acetate is largelyprecipitated from the reaction mixture. After 16 hours, the mixture isfiltered, washed with ether and dried in air.

37.4 grams of 3a-hydroxy-16a,17a-methyleneazo-1l, ZO-pregnanedione3-acetate is placed in a 500 m1. roundbottom flask and heated by an oilbath in vacuo (pressure 0.6 mm.). A manometer and 12 liter surge flaskare in the line between the reaction flask and pump trap. When the bathtemperature reaches 180 C. the 3a-hydroxy 16oc,17oc methyleneazo 11,20pregnanedione 3-acetate begins to melt with evolution of nitrogen. Themaximum pressure reached is 83 mm. After 10 min utes at ISO-182 C. themelt is cooled. It has hgg 249, E percent 191 and is taken up in aboutml. of acetone, filtered through diatomaceous earth, concentrated toabout 100 ml. and ether is slowly added to the boiling solution untilcrystallization occurs. These crystals of SOL-hydroxy l6 methyl 16pregnene 11,20 dione 3- acetate have a melting point of -168" C.

A solution of 20.0 g. of 3u-hydroxy-16-methyl-16- pregnene-11,20-dione3-acetate dissolved in 600 ml. of methanol, is cooled to 18 C., and 80ml. of 30% hydrogen peroxide followed by 80 ml. of 2.5 N sodiumhydroxide are added. Considerable material precipitates from solution,but all redissolves on stirring the reaction mixture at 25 30 C. for 40minutes. The solution is kept at 15 -20 C. for 18 hours at which timethe ultra-violet maximum at 249 has completely disappeared. Then 600 ml.of saturated salt water is slowly added, the crystalline precipitate isfiltered, washed with water, and dried in air and in vacuum. The16a,l7otepoxy 30c hydroxy 16B methyl 11,20 pregnancdione thus formed hasa melting point of l76l77 C.

Room temperature acetylation gives the 3tx-hydroxy-16a,l7a-epoxy-16/3-methyl 11,20 pregnanedi-one 3-acetate having amelting point of 162-164 C.

To a stirred solution of 5.05 g. of3a-hy'droxy-l6ot,l7aepoxy-16fi-methyl-l1,20-pregnanedione 3-acetate in75 ml. of acetic acid maintained at 10l5 C. is added 25 ml. of cold 15%hydrogen bromide in acetic acid. After 35 minutes at 10l5 C. the mixtureis concentrated to dryness in vacuo (temperature 15 C.) and the residuechromatographed on 200 g. of neutral alumina.

From the 50:50 petroleum ether-benzene to 100% benzene eluates, there isobtained 1.60 g. 3a,17u-dihy droxy-16-methylene 11,20 pregnanedione 3acetate; prisms from acetone-ether MP. l98200 C.

Example 2.-Preparatirm of 21-b1'0m0-3ot,17ct-dil1ydr0txy-16-methylane-lLZO-pregnanedione To a stirred solution of 1.60 grams of3a,l7a-dihydroxy-16-methylene-11,20-pregnanedione (prepared as describedhereinbelow) in 32 ml. of chloroform maintained at 3035 C. is slowlyadded 8 40 mg. of bromine in 48 ml. of chloroform. Uptake of bromine iscomplete within 3 hours. Ether is added and the mixture washed twicewith the cold 5% potassium bicarbonate solution. The organic extract isdried over magnesium sulfate and the residue crystallized fromacetoneether to give 21-br0mo-3ot,17ot-dihydroxy-16-methylene-11,20-pregnanedi-one. MP. 190l98 dec.; [vd +34.

Analysis.Ca1Cd. fOI' C H O Br1 C, I'I, Found: C, 60.31; H, 7.38.

The 30,17oc dihydroxy-l6-methylene-1l,20-pregnanedione used as astarting material in this example is prepared starting with the3a,17a-dihydroxy-16-methylene- 11,20-pregnanedione 3-acetate prepared asdescribed in Example 1 in accordance with the following procedure:

To a stirred solution of 3.00 g. of a 36!,l7oc-dlllYdfOXY-l6-methylene-11,20-pregnanedione 3-acetate in 90 ml. of methanol undernitrogen is added 3.00 g. of potassium hydroxide in 1.5 ml. of water and150 ml. of methanol. After 30 minutes at 25, acetic acid (6 ml.) isadded and the solvents removed in vacuo. The product is isolated bychloroform extraction and is crystallized from ether to yield 3a,17x-dihydroxy-16-methylene-l1,20-pregnanedione MP. 160-162; [och 34.

Analysis.Calcd. for C I-1 9 2 C, 73.29; H, 8.99. Found: C, 72.76; H,8.80.

Example 3.Preparatin of 304,] 701,21 -tril1ya'roxy-l 6-methylene-11,ZO-pregnanerlione 3,21-(liacetate 70 mg. of2l-bromo3a,17u-dihydroxy-16-methylene- 11,20-pregnanedione 3-acetate, 55mg. of potassium io dide and 70 mg. of anhydrous potassium acetate inml. acetone and 0.1 ml. acetic acid are refluxed 2 hours. The mixture isconcentrated to dryness in vacuo and partitioned between ethyl acetateand Water. The ethyl acetate layer is washed with sodium chloridesolution and dried over magnesium sulfate. Crystallization of theproduct from acetone-ether gives pure3u,l7a,2l-trihydroxy-16-methylene-1 l,20pregnanedione 3,21-diacetate.MP. 228230 positive tetrazolium test.

12 Analysi.r.Calculated for C H O C, 67.80; H, 7.88. Found: C, 67.28; H,7.75.

Example 4.Preparari0n of 3a,] 7a,21-frilzydr0xy-16- methylene-11,20-pregnancdi0ne 21 acetate In the same manner as in Example 3,2l-bromo-3a,17adihydroxy-16-methylene-11,20-pregnanedione (1.42 g.)(prepared as described in Example 2) is converted to3a,17a,2l-trihydroxy l6 methylene 11,20 pregnancdione ZI-acetate. (1.28g.) M.P. 172-175. [al +44:

Analysis.Calcd. for C H O C, 68.87; H, 8.18. Found: C, 68.47; H, 8.24.

Example 5 .-Preparatian of 1 7u,21 -dihydr0xy-16-met/zylens-3,11,ZO-pregrzanetrfone 21 -acetate To a solution of 600mg. of 3a,17a,21-trihydroxyl6-methylene-11,20-pregnanedione 21-acetatein 16 ml. of acetic acid is added 284 mg. of sodium dichromate in 13 ml.of acetic acid. After 3 hours at 25 C., 50% saturated aqueous sodiumchloride solution is slowly added. The precipitated product is filtered,washed with Water, dried in air and recrystallized from acetone-ether togive pure 17a,21-dihydroxyl6-rnethylene-3,11,20-pregnanetrione 21-actateMP. 202-210".

Example 6 .Preparation of 4-br0m0-1 7a,21-dihydr0xy- 1 6-methylene-3,11,20-pregnanetrione Example 7.Preparalt'0n of 3-semicarbaz0ne 0f 17ot,21-dihydrotxy 16 methylene 4 pregnene 3,11,20- trz'one 21 -acetate To 580mg. of 4-bromo-17a,2l-dihydroxy-l-methylone-3,11,20-pregnanetrioneZl-acetate in 20 ml. of acetonitrile under nitrogen is added a slurry of300 mg. of semicarbazide hydrochloride and 200 mg. sodium bicarbonate in4 ml. of water. After 2 hours, the acetonitrile is removed in vacuo,water added and the crystalline 3-semicarbazone of17a,2l-dihydroxy-l6-methylene- 4-pregnene-3,11,20-trione 2l-acetate isfiltered, Washed with water, and dried in air.

Example 8 .Preparation of 1 7 0:,21 -dihydr0xy-16-methylene-4-pregnene-3,11,20-zri0ne 21 -acetate (16-methylenecortisone acetate) 500 mg. of the semicarbazone of 17a,21-dihydroxy16-methylene-4-pregnene-3,11,20-tri-one is dissolved in 20 ml. of aceticacid, 1.5 ml. of pyruvic acid and 5 ml. of water. After 18 hours at 25C., water is added and the mixture is extracted with chloroform. Thechloroform extract is washed with aqueous potassium bicarbonate, water,and dried over sodium sulfate. Removal of the solvent gives crude17ct,2l-dihydroxy-l6-methylene-4-pregnene-3,l1,20-trione 2l-acetate,which is purified by crystallization from acetone-ether.

Example 9.Preparati0rz of 17u,2l -di/tydr0xy-16-methylene-4-pregnene-3,11,20-tri0ne (16-methylene cortisone) Thiscompound is obtained by treatment of 500 mg. of the corresponding21-acetate (the product of Example 8) in 15 ml. of methanol with 500 mg.of potassium bicarbonate in 5 ml. of water under nitrogen at reflux for10 minutes. The mixture is cooled, neutralized with 0.5 ml. of aceticacid in 5 ml. of water, the methanol removed in vacuo and the productextracted into ethyl acetate. Concentration of the ethyl acetate givescrystalline 17a,2l-dihydroxy-16methylene-4-pregnene-3 ,11,20 trione.

Example J0.Preparation of 3,20 disemz'carbazone of 17a,21 dihydroxy I6methylene 4 pregnene- 3,1 LZO-trione To a stirred solution of 500 mg. of17a,21-dihydroxy- 16-methylene-4-pregnene-3,11,20-trione in 12.5 ml. ofmethanol and 3 ml. of dimethylformamide kept under nitrogen is added aslurry of 680 mg. of semicarbazide hydrochloride and 370 mg. of sodiumbicarbonate in 1 ml. of water. The stirred mixture is refluxed 3 /2hours and maintained at 45 C. for 17 hours. It is then cooled to 20 C.and 50 ml. of 50% saturated aqueous sodium chloride is added. After 2hours at C. the precipitate of the 3,20-disemicarbazone of17u,2l-dihydroxy-16- methylene-4-pregnene-3,11,20-trione ZI-acetate isfiltered, washed with water until free of chloride ion and dried in air.

Example 11.Preparati0n of the 3,20-dz'semicarbaz0ne of ]1B,I7a,21trihydroxy J6 methylene 4 pregnene-3,20-di0ne To a stirred solution of600 mg. of the 3,20-disemicarbazone of17a,2l-dihydroxy-16-methylene-4-pregnene- 3,11,20-trione 21-acetate in30 ml. of tetrahydrofuran and 11 ml. of water under nitrogen is added200 mg. powdered sodium borohydride. The stirred suspension is refluxed45 minutes and then cooled to 15 C. Aqueous acetic acid (3 ml. of 30%)is added cautiously and most of the tetrahydrofuran is removed invacuum. Addition of ml. of methanol and 5 ml. of water induces theproduct to crystallize. Following addition of 10 ml. of a saturatedsodium chloride solution and aging at 0 C. the product of3,20-desimicarbazone of11B,17a,21-trihydroxy-16-methylene-4-pregnene-3,ZO-dione is filtered,washed with water, and dried in air.

Example J2.Preparati0n of 115,17a,2]-trihydr0xy-16- methylene 4 pregnene3,20 dione (16-metlzylene hydrocortz'sone) Example 13.Preparation 0f11/3,]7a,21-trihydr0xy-16- methylene-4-pregnene-3,20-di0ne ZI-acetate(I6-methylene hydrocortisone acetate) The product of the previousExample 12 is acetylated at C-Zl as follows: A solution of 100 mg. of11/3,17oz,21 trihydroxy-l6-methylen-e-4-pregnene-3,20-dione in 1.0 ml.of pyridine and 0.5 m1. of acetic anhydride is prepared. After 18 hoursat 25 C., the solution is taken to dryness in vacuo and the solidresidue purified by crystallization in acetone-ether to givellfi,l7a,2l-trihydroxy- 16-methylene-4-pregnene-3,20-dione 2l-acetate.

Example 14.Preparati0n of 17a,21-dihydr0xy-]6-met/1ylene-J,4-pregnadiene-3,1l,20-tri0ne Treatment of 17:1,21 dihydroxy16-methylene-4-pregnene-3,11,20-trione prepared as in Example 9 or thecorresponding 21-acetate prepared as in Example 8 with Bacillussplzaericus under suitable conditions leads to 17a,

7 21 dihydroxy-16-methylene-1,4-pregnadiene-3,11,20-tri- ODE.

1 4 Example 15.Preparati0nof 2,4-dibr0m0-1 711,21 -dihydroxy-l6-methylene-3,11 ,ZO-pregnanetrione 21 -acetate To a stirred solution of200 mg. of Nail-dihydroxy- 16-methylene-3,11,20-pregnanetrione2l-acetate (prepared as described in Example 5) in 4 ml. of chloroformand 0.05 ml. of acetic acid maintained at 15 C. is added dropwiseone-half of a solution of 162 mg. of bromine in 0.47 ml. of chloroformand 0.53 ml. of acetic acid. The mixture is warmed to 0 C. and theremainder of the bromine added. Sodium acetate (0.1 g.) in 0.4 ml. ofwater is added followed by a few drops of 5% aqueous sodium sulfite.Additional chloroform and water are added and the mixture is extractedwith chloroform. The chloroform layer is washed with aqueous potassiumbicarbonate, aqueous sodium chloride and dried over magnesium sulfate.Evaporation of the solvent leaves as a residue 275 mg. of2,4-dibromo-17a,21-dihydroxy- 16-methylene-3,11,20-pregnanetrione21-acetate.

Example 16.-Preparati0n 0 170:,21 dihydroxy 16-methylene-1,4-pregnadiene-3,11,20-tri0ne 21 -acetate CHaOH max.

238 mp. Em 14,400; [a] -l123 Analysis.Calculated for C H O C, 69.88; H,6.84. Found: C, 69.96; H, 6.94.

Example 17.Preparation of 17a,21 dihydroxy 16- methylene-1,4-pWegnadiene-3J 1 ,20-tri one This compound is obtained by treatment of1.0 g. of 17a,21-dihydroxy 16-methylene-1,4-pregnadiene-3,11,20- trioneZI-acetate in 30 ml. of methanol with 1 g. of potassium bicarbonate in10 ml. of water under nitrogen at reflux temperature for 7 minutes. Themixture is cooled, neutralized with 1 ml. of acetic acid in 10 ml. ofwater, the methanol removed in vacuo and the product is extracted intoethyl acetate. Removal of the ethyl acetate gives 170:,21dihydroxy-16-methylene-1,4-pregnadiene-3, 1 1,20-trione.

Example 18.Preparati0n 0f 11/3,17a,21 trihydroxy- I6-methylene-1,4-pregnadiene-3,20-dione 21 -acetate The1,2-dehydrogenation of 11,8,17a,21-trihydroxy-16-methylene-4-pregnene-3,20-dione 21-acetate prepared as in Example 13 maybe carried out microbiologically by means of Bacillus sphaericus toyield 11fl,17u,21-trihydroxy-16-methylene-1,4-pregnadiene-3,20-dione,which on acetylation with acetic anhydride-pyridine gives 115,17a,

21 trihydroxy-16-methylene-1,4-pregnadiene-3,20-dione ZI-acetate.

Example 19.-Preparation 0 the 3,20 disemicarbazone of a fluoro11/3,]7a,21-trihydr0xy-16a-methyl-L4- p-regnadiene-3,20-dione A mixtureof 1.00 g. of9a-fluoro-11/3,17a,21-trihydroxy-16a-met-hyl-1,4-pregnadiene-3,ZO-dione,750 mg. of semicarbazide base, 280 mg. of semicarbazide hydrochloride in20 ml. of methanol and 10 ml. of dimethylformarnide is refluxed for 20hours under nitrogen. The mixture is cooled to 20 C. and ml. of water isadded with stirring. The precipitated 3,20-disemicarbazone of 9:1fluoro-11B,17a,2l-trihydroxy-16u-methyl-1,4-pregnadiene-3,20-dione isfiltered, washed with water, and dried in air; M.P. over 300;

In similar manner are prepared the 3,20-disemicarbazone of 9ot-fluoro-11B,17e,21-trihydroxy-16,8-methyl-1, 4-pregnadiene-3,20-dione,3,20-disemicarbazone of 11,8, 1701,21 trihydroxy16umethyl-1,4-pregnadiene-3,20-dione; 3,20 disemicarbazone of 11B,l7a,21tri-hydroxy- 16i3-methyl-1,4-pregnadiene-3,20 dione; 3,20disemicarbazone of Hall-dihydroxy-la-methyl-l,4-pregnadiene-3,11,20-trione; 3,20-disemicarbazone of flail-dihydroxy- 16,8-methyl1,4-pregnadiene-3,11,20-trione; 3,20-disemicarbazone of Qoc-fillOYO-l13,1701,21-trihydroxy-l6a-methyl- 4-pregnene-3,20-dione;3,20-disemicarbazone of 9a-fluoro- 11,8,17u,21 trihydroxy 16Bmethyl-4-pregnene-3,20 dione; 3,20 disemicarbazone of 11fi,17oc,21trihydroxy- 161x methyl 4 pregnene-3,20 dione; 3,20-disemicarbazone of11,8,17o,21 trihydroxy 165-methyl-4-pregnene-3,20-dione; 3,20disemicarbazone of 17a,21-dihydroxy-16a methyl 4 pregnene 3,11,20-trioneand 3,20 disemicarbazone of 1711,21 dihydroxy-16B-methyl- 4-pregnene-3,11,20-trione.

Similarly, starting with 9a-chloro and 9a-bromo derivatives of11,3,17u,21 trihydroxy-I6u-methy1-L4-pregnadiene-3,20-dione, there areobtained the 3,20-disemicarbazone of 90a chloro 115,17et,21 trihydroxy16emethyl 1,4 pregnadiene 3,20-dione and the 3,20-disemicarbazone of 90abromo 11B,170c,21 trihydroxy- 16a methyl-1,4 pregnadiene 3,20 dione; andstarting with the 901 chloro and 9a bromo derivatives of 11B,17a,21trihydroxy 16a methyl 4 pregnene-3,20- dione, there are obtained the3,20 disemicarbazone of 9a chloro 11B,17ot,21 trihydroxy 16a methyl-4-pregnene 3,20 dione and the 3,20 disemicarbazone of 90: brorno1lfi,l7x,21 trihydroxy 16a-methyl-4- pregnene-3,20-dione.

Example 20.Preparatin of the 3,20-disemicarbaz0ne 0f 9 fluoro-1701,21dihydroxy 16a methyl-1,4-pregnadime-3,11 ,ZO-trione A mixture of 1.00grams of 90a fluoro 170c,21 dihydroxy 161x methyl 1,4pregnadiene-3,11,20-trione, 750 mg. of semicarbazide base, 280 mg. ofsemicarbazide hydrochloride in 20 ml. of methanol and ml. ofdirnethylformamide is refluxed for 20' hours under nitrogen. The mixtureis cooled to 20 C. and 100 ml. of water is added with stirring. Theprecipitated 3,20-disemicarbazone of9a-f1uoro-17a,21-dihydroxy-16a-methyll,4-pregnadiene-3,11,20-trione isfiltered, washed with Water, and dried in air.

Similarly, starting with the 9oc-Chl01'0 and 9u-bromo derivatives of1701,21 dihydroxy 16oz methyl 1,4- pregnadiene-3,l1,20-trione, there areobtained the 3,20- disemicarbazone of 9a chloro17a,21-dihydroxy16ocmethyl-1,4-pregnadiene-3,11,20-trione and the3,20-disemicarbazone of 9a brorno17a,21-dihydroxy-16amethyl-1,4-pregnadiene-3 ,1 1,20-trione.

In accordance with the foregoing procedure, but starting with 90: fluorol7a,21-dihydroxy-16a-methyl-4- pregnene-3,l1,20-trione, there isobtained the 3,20-d'isemica-rbazone of 90a fluoro 170:,21dihydroxy-16a-methyl- 4-pregnene-3,11,20-trione.

Similarly, starting with the 9ot-chloro and 9a-bromo derivatives of17u,21-dihydroxy-16a-methyl-4-pregnene- 3,11,20-trione, there areobtained the 3,20-disemicarbazone of 9oz chloro 1701,21 dihydroxy 16amethyl- 4-pregnene-3,11,20-trione and the 3,20-disemicarbazone of 90:bromo 17a,21 dihydroxy 16a methyl 4- p s L O-tI Qne 1% Example21.Preparatiorz of 9a fluoro 115,21 [Iihydroxy 16 methyl 1,4,16pregnatriene 3,20- dione 21 acetate A solution of 500 mg. of the3,20-disemicar-bazone of diene-3,20-dione in 10 ml. of acetic acid and0.5 ml. acetic anhydride is refluxed under nitrogen for one hour toproduce the corresponding 3,20-disemicarbazone of 9a fluoro115,2l-dihydroxy-l6-methyl-1,4,16-pregnatriene-3,20-dione 21-acetate.The reaction mixture is cooled, 13 ml. of Water is added and the mixtureheated on the steam bath for 5 hours. It is then concentrated in vacuonearly to dryness and water and chloroform added. The mixture isthoroughly extracted with chloroform, and the chloroform extract Washedwith excess aqueous potassium bicarbonate, saturated salt solution anddried over magnesium sulfate. Chromatography of the residue on neutralalumina and crystallization of pertinent benzene-chloroform fractionsgives 901-111101'0-1113, 21-dihydroxy 16 methyl 1,4,16pregnatriene-3,20- dione ZI-acetate; M.P. 228-233;

n 243 m Em 22,000

Similar treatment of the 3,20-disem'icarbazone of 9afluoro 1l13,l7a,21trihydroxy 16,8 methyl 1,4- pregnadiene 3,20 dione also leads to9u-fluoro-11fi,21- dihydroxy 16 methyl 1,4,6 pregnatriene 3,20-dione21-acetate.

In a similar manner 115,21 dihydroxy-16-methyl- 1,4,l6-pregnatriene-3,ZO-dione Ill-acetate; 21-hydroxy-l6- methyl 1,4,16pregn-atriene 3,11,20-trione 21-acetate; 9a fluoro 115,21 dihydroxy 16methyl 4,16- pregnadiene-Ii,20-dione 2 l-acetate; 1 113,21-dihydroxy-16-methyl 4,16 pregnadiene 3,20-dione 21-acetate; and 21-hydroxy 16 methyl4,16 pregnadiene-3,11,20- trione 21-acetate are obtained.

Similarly, starting with the 9a-chloro and 9a-bromo derivatives of the3,20-disemicarbazone of 11/3,17u,21- trihydroxy 16a methyl 1,4pregnadiene 3,20dione, there are obtained the 91x chloro 116,21dihydroxy- 16 methyl 1,4,16 pregnatriene 3,20 dione 21-acetate and abrorno 11,8,21 dihydroxy16-rnethyl-l,4,16- pregnatriene 3,20 dione21-acetate; and starting with the 9a-chloro and 9a-bromo derivatives ofthe 3,20-disemicarbazone of 11fi,17a,21-trihydroxy 16a methyl-4-pregnene-3,20-dione, there are obtained 9u-ChlOIO-11B, 21 dihydroxy 16methyl 4,16 pregnadiene-3,20- dione 2l-acetate and 9m brorno 115,21dihydroxy- 16-methyl-4,16-pregnadiene-3 ,20-dione 21-acetate.

Example 22.Preparation of 9-flu0r0-21-hydr0xy-16-methyl-1,4,I6-pregnatriene-3,11,20-tri0ne 21-acetate A solution of 500mg. of the 3,20-disemicarbazone of 90a fluoro 1701,21dihydroxy-16a-methyl-1,4-pregnadiene-3,11,20-trione in 10 ml. of aceticacid and 0.5 ml. acetic anhydride is refluxed under nitrogen for onehour to produce the corresponding 3,20-disemicarbazone of 9a fluoro 21hydroxy 16-methyl-1,4,16-pregnatriene- 3,11,20-trione 21-acetate Thereaction mixture is cooled, 13 ml. of water is added and the mixtureheated on the steam bath for 5 hours. It is then concentrated in vacuonearly to dryness and Water and chloroform added. The mixture isthoroughly extracted with chloroform, and the chloroform extract Washedwith excess aqueous potassium bicarbonate, saturated salt solution anddried over mag nesium sulfate. Chromatography of the residue on neutralalumina and crystallization of pertinent benzenechloroform fractionsgives 9a-fluoro 21 dihydroxy- 16-methyl1,4,16-pregnat-riene-3,11,20-trione 2l-acetate; M.P. 228233;

x11 3 243 m Em 22,000

Similarly, starting with the 9a-chloro and 9a-bromo derivatives of the3,20-disemicarbazone of 17 u,21-dihydroxy 16a methyl 1,4 pregnadiene3,11,20-trione,

there are obtained 90: chloro 21 hydroxy 16 methyl- 1,4,16 pregnatriene3,11,20-trione 21-acetate and 9c:- bromo 21 hydroxy 16 methyl1,4,16-pregnatriene- 3,11,20-trione 2l-acetate.

In accordance with the foregoing procedures, but starting with the3,20-disemicarbazone of 9a-fluoro-17a,21- dihydroxy 16cc methyl 4pregnene 3,11,20-trione, there is obtained 9a fluoro 21 hydroxy 16methyl- 4,l6-pregnadiene-3,11,20-trione 21-acetate.

Similarly, starting with the 9oc-Chl0l0 and 90C-b'I'OIIlO derivatives ofthe 3,20-disemicarbazone of 17a,21-dihydroxy 160a methyl 4 pregnene3,11,20-trione, there are obtained 90c chloro 21 hydroxy 16 methyl-4,l6-pregnadiene-3,l1,20-trione 21-acetate and 9a-bromo-21-hy-droxy-16-methyl-4,l6-pregnadiene-3 ,1 1,20-trione 21- acetate.

Example 23.Preparatin 0f 9u-flu0r0-1I5,21-dilzydr0xy- I65methyl-16a,17a-0xid0-1,4-pi'egnarliene-3,20-d[0ne 21 -acetate To astirred solution of 500 mg. of 9u-fiuoro-l15,21-dihydroxy-l6-rnethyl-l,4,16-pregnatriene-3,20 dione 21- acetate in 5 ml.of t-butyl hydroperoxide and 0.1 ml. of a methanolic solution ofbenzyl-trimethyl ammonium hydroxide. After 18 hours at room temperature,water is added and the mixture thoroughly extracted with chloroform. Thechloroform extract is washed with saturated aqueous sodium chloride anddried over magnesium sulfate. Evaporation of the solvent andcrystallization of the residue from acetone-ether gives 9a-fiuoro-115,21- dihydroxy 165 methyl-16a,l7a-oxido-1,4-pregnadiene- 3,20-dione 21-acetate.

In a similar manner are prepared l15,2l-dihydroxy-165-methyl-1611,17a-oxido-1,4-pregnadiene-3,ZO-dione 21- acetate; 21hydroxy-165-methy1-16a,17a-oxido-1,4-pregnadiene-3,ll,20-trione21-acetate; 9a-fiuoro-115,21-dihydroxy 165methyl-l6a,17a-oxido-4-pregnene-3,20-dione 21-acetate;115,21-dihydroxy-165-methyl-16a,17ot-oxido- 4-pregnene-3,20-dione21-acetate, and 21-hydroxy-165- methyl-l61,l7a-oXidopregnene 3,11,20trione 21- acetate.

Similarly, starting with the 9u-Cl1l010 and 9m-bromo derivatives of l15,2l-dihydroxy-l6-methyl-1,4,16-pregnatriene-3,20-dione 21-acetate,there are obtained 9achloro1l5,2l'dihydroxy-165-methyl-16a,17a-oxido-1,4- pregnadiene-3,20-dione21-acetate and 9ot-bromo-115,21- dihydroxy 165methyl-16oz,17a-oxido-1,4-pregnadiene- 3,20-dione Zl-acetate; andstarting with the 9a-chloro and 9ot-bromo derivatives of115,21-dihydroxy-16-methyl- 4,16-pregnadiene-3,20-dione 21-acetate,there are obtained 9a chloro l15,2l-dihydroxy-165-methyl-16a,l7ct-oxido-4-pregnene-3,20-dione 2l-acetate and 9a-hromo-1l5,21- dihydroxy 165methyl-16a,17a-oxido-4-pregnene-3,20- dione 21-acetate.

To a stirred solution of 500 mg. of9a-fluoro-21-hydroxy-16-methy1-1,4,16-pregnatriene 3,11,20 trione 21-a-cetate in 5 ml. of benzene and 5 ml. of chloroform are added 0.50 ml.of t-butyl hydroperoxide and 0.1 ml. of a 35% methanolic solution ofbenzyl-trimethyl ammonium hydroxide. After 18 hours at room temperature,water is added and the mixture thoroughly extracted with chloroform. Thechloroform extract is Washed with satu rated aqueous sodium chloride anddried over magnesium sulfate. Evaporation of the solvent andcrystallization of the residue from acetone-ether gives 9a-fluoro-21-hy-(lI'OXY-16/3-Hl6tl'lYl-160t,170t-OXlClOrl,4 pregnadiene 3,11, ZO-trione21-acetate.

Similarly, starting with the 9oz-Chl010 and 9OC-bl'OIT1O derivatives of2l-hydroxy-l6-methyl-l,4,16-pregnatriene- 3,11,20-trione 21-acetate,there are obtained 9a-chloro- 21 hydroxy165-methyl-l6a,l7u-oxido-1,4-pregnadiene- 3,11,20-trione 2l-acetate and9a-bromo-21-hydroxy-165- Example 25.-Preparali0n ofQa-fluorO-II/i,l7a,21-trihydroxy-I6-methylene-1,4-pregnadiene 3,20 dione21- acetate To a stirred solution of 600 mg. of9ot-fluoro-1l5,2ldihydroxy methyl-l6a,17a-oxido-l,4-pregnadiene-3,20-dione 21-acetate in 10 ml. of acetic acid maintained at 10-15 C. isadded 3 ml. of cold 10% hydrogen bromide in acetic acid. After 30minutes the mixture is concentrated to dryness in vacuo (temp. 15) andthe residue chromatographed on neutral alumina. Combination of pertinentbenzene-chloroform fractions and crystallization leads to the desired9a-fluoro-ll5,17a,2l-trihydr0xy-16- methylene-1,4-pregnadiene-3,20-dione21-acetate.

In a similar manner are prepared115,17a,2l-trihydroxy-l6-methylene-l,4-pregnadiene-3,20 dione2l-acetate; :,21 dihydroxy-l6-methylene-1,4-pregnadiene-3, 11,20-trioneZI-acetate; 9a-fiuoro-115,17a,21-trihydroxy-16-methylene-4-pregnene-3,20-dione ZI-acetate; 115,17a,2l-trihydroxy-l6-methylene-4-pregnene 3,20 dione 21- acetate; and17:1,21-dihydroxy-l6-methylene-4-pregnene- 3,1 1,20-trione 21-acetate.

Alternatively, to a stirred solution of 600 mg. of 9afiuoro 115,21dihydroxy-165-methyl-16a,l7a-oxido-1,4- pregnadiene-3,20-dione2l-acetate in 2.4 ml. of tetrahydrofuran is added 3.5 m1. of a 2:1 byweight mixture of hydrogen fluoride in tetrahydro-furan. After 2 hoursat 15 the reaction mixture is pipetted slowly into excess cold aqueoussodium carbonate solution. The mixture is extracted with chloroform andthe chloroform extract washed with water and saturated aqueous sodiumchloride. Chromatography of the residue on neutral alumina as above alsoleads to the desired9a-flu0ro-l15,17u,2l-trihydroxy-l6-methylene-1,4-pregnadiene 3,20 dione21- acetate.

Similarly, starting with the 9a-chloro and 9a-bromo derivatives of115.21 dihydroxy 165 methyl-16a,l7aoxido-l,4-pregnadien-e-3,20-dione21-a'cetate, there are obtained9a-chloro-115-1704,21-trihydroxy-16-methylene-1,4-pregnadiene-3,20-dione 21-acetate and 9a-bromo-115-17ot,21-trihydroxy-16-methylene-1,4-pregnadiene 3,20 dione ZI-acetate; andstarting with the 9oc-Chl010 and 9a-bromo derivatives of 115,21dihydroxy 165 methyl-16a,17 xoxido-4-pregnene-3,20-dione 21-acetate,there are ob.- tained 90 chloro-l15,17a,21-trihydroxy-16-methylene-4-pregnene-3,20-dione 21-acetate and 9a-bromo-ll5,l7a,21-trihydroxy-16-methylene-4-pregnene 3,20 dione 2l-acetate.

Example 26.Preparati0rz of 9a flu0ro-1 7a,21-dihydr0xy- 16-metlzylene-I,4-pregnadiene-3,11,20-tri0ne 21 acezate To a stirredsolution of 600 mg. of 9a-fluoro-21-hydroxy-l65-methyl-16a,17a-oxido 1,4pregnadiene-3,l1, 20-trione 2l-acetate in 10 ml. of acetic acidmaintained at 10-15 C. is added 3 ml. of cold 10% hydrogen bromide inacetic acid. After 30 minutes the mixture is concentrated to dryness invacuo (temp. 15) and the residue chromatographed on neutral alumina.Cornbina tion of pertinent benzene-chloroform fractions andcrystallization leads to the desired 9a-fiuoro-17u,21-dihy- I3,11,20-trione 21-acetate.

Similarly, starting with the 90t-ChlOI'O and 9a-bromo de rivatives of21-hydroxy-16 3-met-hyl-16a,l7ot-oxido-4-pregnone-3,11,20-trione2l-acetate, there are obtained 90:- chloro l7oc,2l dihydroxy 16methylene 4 pregnene- 3,11,20-trione Zl-acetate and9cx-bf0m0-170:,21-dlhYClf0XY- l6-methylene-4-pregnene-3,11,20-trione21-acetate.

This compound is obtained by treatment of 200 mg. of the corresponding21-acetate (the product of Example 22) in 10 ml. of methanol with 200mg. of potassium bicarbonate in 3 ml. of water under nitrogen at refluxfor 10 minutes. The mixture is cooled, neutralized with 0.3 ml. ofacetic acid in 5 ml. of Water, the methanol removed in vacuo and theproduct extracted into ethyl acetate. Concentration of the ethyl acetategives crystalline 9a-fiuoro-l 1e, 17a,2l-trihydroxy-16-methylene-1,4-pregnadiene-3,20-dione.

In a similar manner may be obtained ll/3,l7a,2l-trihydroxy l6methylene-l,4-pregnadiene-3,ZO-dione; 17a, 21-dihydroxy-l6-methylene 1,4pregnadiene 3,11,20- trione; 9a fiuoro-l13,17a,2l-trihydroxy-16methylene-4- pregnene-3,20-dione;11,8,17a,21-trihydroxy-1-methylene- 4-pregnane-3,20-dione andl7a,2l-dihydroxy-l6-rnethylene-4-pregnene-3,l1,20-trione.

Emmple 28.-Preparati0n of Qcc-flLlOi'O-I 16,1 70,2l -trihydroxy-l(i-methylene] ,4-pregnadiene-3,20-dione 100 mg. of9a-fiuoro-l1,5,17at,21-trihydroxy-l6-methylene-4-pregnene-3,20-dione21-acetate is prepared as described in Example 22, and is treated withBacillus Splittericus to produce the corresponding 9a-fi .\oro-ll,8,17t,2ltrihydroxy-l6-methylene-l,4-pregnadiene-3,ZO-dione.

Example 29.-Preparati0n of 9w hero-118,17a,21-trilzydroxy 16merhylane-1,4-pregnadiene3,20-di0ne 21-1- butyI-acetate To a stirredsolution of 50 mg. of 9ot-fiuoro-11fl,17a,21-trihydroxy-l6-n1ethylene-1,4-pregnadiene-3 ,ZG-dione (the product of theprevious example) in 0.5 ml. of pyridine at is added 0.3 ml. oft-butylacetyl chloride. After two hours at 25 Water is added and theprecipitated product filtered, washed with dilute hydrochloric acid,dilute sodium bicarbonate solution, water and dried in air.

In a similar manner may he obtained the ZI-t-butyl acetates of l18,l7a,2l-trihydroxy-16-methylene-1,4-pregnadiene-3,20-dione; 1741,21dihydroxy-l6-methylene-1,4- pregnadiene 3,11,20 trione;9wfiuoro-l1,8,17a,21-trihydroxy 16 methylene-4-pregnene-3,ZO-dione;11B,17oc,21- trihydroxy-l6-methylene-4-pregnene-3,ZO-dione and 170:,21-dihydroxy-1 6-methylene-4-pregnene-3 ,1 1,20-trione.

In an analogous manner may be prepared the corresponding 2l-propionates2l-benzoates and other 21-esters of aliphatic and aromatic acids.

Example 30.Preparati0rz of 90z-flll0iO-17a,21dll1ydl'0,\IG-methylane-,4-pregnadiene-3,11,20-tri0ne 21-acetate To a stirredsolution of 120 mg. of Qot-flUOIO-llfi,l7ot,2l-trihydroxy-16-methylene-1,4-pregnadiene 3,20 dione ZI-acetate in 3ml. of glacial acetic acid is added 43 mg. of sodium dichrornate in 2ml. of glacial acetic acid at 20? C. The mixture is stirred at 20'25 for3 hours,

diluted with water and extracted with chloroform. The chloroform extractis washed with aqueous potassium bicarbonate solution, saturated sodiumchloride solution, dried over magnesium sulfate and taken to dryness.Crystallization of the residue from acetone-ether givesiluoro-17rx,21-dihydroxy-16-methylene 1,4 pregnadiene- 3,11,20-trione21-acetate. The corresponding 2l-alcohol is obtained by treatment of the21-acetate with aqueous methanolic potassium bicarbonate as described inExample 17.

In a similar manner 9a-fluoro-l7u,2l-dihydroxy-l6methylene-4-pregnene-3,11,20-trione ZI-acetate and 21- alcohol areobtained from 9a-fluoro-11fl,17a,2l-trihydroxy- 16-n1ethylene-4-pregnene-3 ,20-dione 21-acetate.

Various changes and modifications may be made in the present invention,certain preferred embodiments of which are herein disclosed, withoutdeparting from the scope thereof; to the extent that these changes andmodifications are Within the scope of the appended claims, they are tobe considered a part of this invention.

Example 31 A solution of 3.0 grams of17a,21-dihydroxy-16-methylene-4-pregnene-3,l1,20-trione Zl-acetate in 20ml. of methanol is reduced in hydrogen at 1 atmosphere and 25 C. in thepresence of 2.0 grams of 25% palladiumon-calcium carbonate catalyst.Hydrogen uptake is complete in 45 minutes. The mixture is filtered, thefiltrate is taken to dryness and the residue crystallized from ether.Chromatography of 1 gram of the product on grams of activated magnesiumsilicate resolves the mixture into l7e,2l-dihydroxyfi6a-methyl-pregnane-3,11,20-trione 21-acetate and 17a,2ldihydroxy-165-methyl-pregnane-3,11,20-trione 21-acetate.

In accordance with the foregoing procedures, but starting with115,170:2l-trihydroxy-16-methylene-1,4- pregnadiene-3,20-dione 21acetate, there are obtained the corresponding 1lfi,l7a,2ltrihydroxy-16a-rnethyl-pregnane- 3,20-dione 21-acetate and1155170;,21-trihydroxy-16flmethyi-pregnanc-S,ZO-dione Zl-acetate.

In accordance with the foregoing procedures, but start ing with9ot-fluoro-17m,21-dihydroxy-16-methylene-4- pregnene-3,1l,20-trione2l-acetate, there are obtained the corresponding 90: tluoro17a,2l-dihydroxy-l6amethyl-pregnane-3,l1,20-trione 21-acetate and9a-fiu01'0 17a,21 dihydroxy 16flmethyl-pregnane-3,l1,20-trioneZI-acetate.

In accordance with the foregoing procedures, but startin g with9a-fl110l'0-1 1B,17a,2l-trihydroxy-16-methylene-1,4-pregnadiene-3,ZO-dione ill-acetate, there are obtained thecorresponding9a-fiuor0-11(3,17a,21-trihydroxy-l6umethyl-pregnane-3,ZO-dione21-acetate and 9a-fluoro- 11,B,17oc,2ltrihydroxy-16fi-methyl-pregnane-3,ZO-dione ZI-acetate.

In accordance with the foregoing procedures, but starting with 17a,2ldihydroxy-16-1nethylene-3,11,20- pregnane-trione ZI-acetate, there areobtained the corresponding 17a,2l-dihydroxy-l6ot-methyl-pregnane-3,l1,20-trione 21-acetate and 17,2-l dihydroxy-l6[3-methy1- pregnane-3,11,20-trione 21-acetate.

in accordance with the foregoing procedures, but starting with11B,17e,21-trihydroxy-16 methylene-3,20-pregnane-dione ZI-acetate, thereare obtained the corresponding 11B,17o ,21 trihydroxy16ot-methyl-3,20-pregnanedione 21-acetate and1lfi,17u,21-trihydroxy-l6u-methyl- 3,2(hpregnane-dione 21-acetate.

Example 32 2.1 chloroform extract is washed with dilute potassiumbicarbonate, water, and dried over sodium sulfate. The residue istriturated with ether to give crystalline 4-bromo- 17a,21 dihydroxy16a-methyl-pregnane-3,11,20-trione 2l-acetate.

To 580 mg. of 4-bromo-17a,21-dihydroxy-16a-methylpregnane-3,11,20-trioneZI-acetate in 20 ml. of acetonitrile under nitrogen is added a slurry of300 mg. of semicarbazide hydrochloride and 200 mg. sodium bicarbonate in4 ml. of water. After 2 hours the acetonitrile is removed in vacuo,water added and the crystalline 3-semicarbazone of 17a,21dihydroxy-16a-methyl-4-pregnene- 3,11,20-trione 21-acetate is filtered,washed with water and dried in air.

500 mg. of the 3-semicarbazone of Wall-dihydroxy-16a-methyl-4-pregnene-3,l1,20-trione 21-acetate is dissolved in 20 ml.of acetic acid, 1.5 ml. of pyruvic acid and 5 ml. of water. After 18hours at 25 C., water is added and the mixture is extracted withchloroform. The chloroform extract is washed with aqueous potassuimbicarbonate, water, and dried over sodium sulfate. Removal of thesolvent gives crude 17a,21-dihydroxy-16amethyl-4-pregnene-3,11,20-trioneZI-acetate, which is purified by crystallization from acetone-ether.

This product is then hydrolyzed by treatment with a solution ofpotassium bicarbonate in aqueous methanol to form 17zx,21 dihydroxy16a-methyl-4-pregnene- 3,11,20-trione.

In accordance with the foregoing procedures, but starting with 11B,17u,2l-trihydroxy-1 6a-methyl-pregnene- 3,20-dione 21-acetate, thereare obtained the corresponding 1118,1704,21-trihydroxy-16a-methyl-4-pregnene-3,20- dione and the 21-acetatethereof.

In accordance with the foregoing procedures, but starting with1lfi,17a,21-trihydroxy-16fi-methyl-pregnene- 3,20-dione ZI-acetate,there are obtained the corresponding 11,8,17a,21trihydroxy-16,3-methyl-4-pregnene-3,20- dione and the 21-acetatethereof.

In accordance with the foregoing procedures, but starting with 90;fluoro 17a,21-dihydroxy-16a-methylpregnane-3,11,2 -trione 21-acetate,there are obtained the corresponding 9a fiuoro17a,21-dihydroxy-16umethyl 4 pregnene-3,11,20-trione and the 21-acetatethereof.

In accordance with the foregoing procedures, but starting with 9:1fluoro 11,8,17a,21-trihydroxy-16fimethyl-pregnane-Ia,20-dione21-acetate, there are obtained the corresponding9a-fluoro-l15,l7a,2l-trihydroxy-16B- methyl-4-pregnene-3,20-dione andthe 21-acetate thereof.

Example 33 To a stirred solution of 200 mg. of 17a,2l-dihydroxy-16a-methyl-pregnane-3J1,20-trione 21-acetate in 4 ml. of chloroform and0.05 ml. of acetic 'acid maintained at 15 C. is added dropwise one-halfof a solution of 162 mg. of bromine in 0.47 ml. of chloroform and 0.53ml. of acetic acid. The mixture is warmed to 0 C. and the remainder ofthe bromine added. Sodium acetate (0.1 g.) in 0.4 ml. of water is addedfollowed by .a few drops of aqueous sodium sulfite. Additionalchloroform and water are added and the mixture is extracted withchloroform. The chloroform layer is washed with aqueous potassiumbicarbonate, aqueous sodium chloride and dried over magnesium sulfate.Evaporation of the solvent leaves as a residue 275 mg. of2,4-dibromo-l7a,21-dihydroxy-l6a-methyl-pregnane-3,l1,20-trione21-acetate.

To a solution under nitrogen of 275 mg. of the 2,4-dibromo 1701,21dihydroxy 16oz methyl pregnane 3, 11,20-tri0ne 21-acetate in 3.3 ml.dimethyl formamide is added 53 mg. of sodium bromide. After one hour at22 25 C. 0.66 ml. of dimethyl aniline is added and the mixturemaintained at C. for 2% hours. The mixture is cooled, added dropwise todilute hydrochloric acid and solid, crude product filtered, washed withdilute hydrochloric acid, water and dried in air. Treatment withcharcoal followed by chromatography on neutral alumina gives1704,21-dihydroxy-16a-rnethyl-1,4-pregnadiene-3,11, ZO-trione21-acetate. This product is then hydrolyzed by treatment with 'asolution of potassium bicarbonate in aqueous methanol to form17ot,21-dihydroxy-16a-methyll,4-pregnadiene-3,1 l,20-trione.

In accordance with the above procedures, but starting with11B,170,2l-trihydroxy-16ct-methyl-pregnane-3,20-dione 21-acetate, thereare obtained the corresponding 115, 1711.21 trihydroxy 16c. methyl 1,4pregnadiene 3, 20-dione and the 21-acetate thereof.

In accordance with the above procedures, but starting with9a-fluor0-17a,2l-dihydroxy-l6ot-methyl-pregnane-3, 11,20-trioneZI-acetate, there are obtained the corresponding 90: fluoro 17a,21dihydroxy 16cc methyl 1,4- pregnadiene-3,l1,20-trione and the 21-acetatethereof.

In accordance with the above procedures, but starting with 904 fiuoro11fl,17a,21 trihydroxy 16B methylpregnane-3,20-dione 21-acetate, thereare obtained the corresponding 90: fluoro 11,8,17a,21 trihydroxymet-hyl-l,4-pregnadiene-3,20-trione and the 21-acetate thereof.

Various changes and modifications may be made in the present invention,certain preferred embodiments of which are herein disclosed, withoutdeparting from the scope thereof; to the extent that these changes andmodifications are within the scope of the appended claims, they are tobe considered a part of this invention.

We claim:

1. A compound selected from the group consisting of:

---o H J CHz I l and the A derivative thereof wherein X is selected fromthe group consisting of hydrogen and tat-halogen, Y is selected from thegroup consisting of keto and fl-hydroxy; 21-position esters and acidesters thereof derived from mono and dicarboxylic acids containing onlycarbon, hydrogen and oxygen up to a total of twenty carbon atoms.

2. A pharmaceutical composition comprising a compound as claimed inclaim 1 together with a pharmaceutically acceptable carrier.

3. 9a fluoro 16 methylene A pregnene 115,170, 21-triol-3,20-dione21-acetate.

4. 9a fluoro 16 methylene A pregnene 1704,21- diol-3,11,20-trione2l-acetate.

5. 9a fluoro 16 methylene A pregnadiene- 1 1,8,17a,21-triol-3,20-dione21-acetate.

6. 9oz fluoro 16 methylene A 17o,21-di0l-3,11,20-trione 21-acetate.

- pregnadiene- No references cited.

ELBERT L. ROBERTS, Primary Examiner. MORRIS LEEMAN, Examiner. LEWISGOTTS, Assistant Examiner.

1. A COMPOUND SELECTED FROM THE GROUP CONSISTING OF:
 2. A PHARMACEUTICALCOMPOSITION COMPRISING A COMPOUND AS CLAIMED IN CLAIM 1 TOGETHER WITH APHARMACEUTICALLY ACCEPTABLE CARRIER.